Abstract

and non-union of bony fractures has been proposed since 1966, little has been known
about the effect of HBOT on bone marrow stem cells (BMSC). The aim of this study
is to investigate the effect of HBO treatment on osteogenetic differentiation of BMSC
and potential application in bone tissue engineering.
Adhesive stromal cells harvested from bone marrow were characterized by
mesenchymal differentiation potential, cell surface markers and their proliferation
capacity. Mesenchymal stem cells, which demonstrated osteogenic, chondrogenic and
adipogenic differentiation potential and expressed positively for CD 29, CD 44, CD
73, CD 90, CD 105, CD 166 and negatively for CD34 and CD 45, were selected and
treated in a laboratory-scale HBO chamber using different oxygen pressures and
exposure times. No obvious effect of HBO treatment on BMSC proliferation was
noticed. However, cytotoxic effects of HBO were considerably less pronounced when
cells were cultured in medium supplemented with 10% FBS in comparison to medium
supplemented with 2% FCS, as was evaluated by WST-1 assay. Under HBO
treatment, bone nodules were formed in three days, which was clearly revealed by
Von Kossa staining. In contrasts, without HBO treatment, bone nodules were not
detected until 9-12 days using the same inducing culture media. Calcium deposition
was also significantly increased after three days of HBO treatments compared to no
HBO treatment. In addition it was also found that oxygen played a direct role in the
enhancement of BMSC osteogenic differentiation, which was independent of the
effect of air pressure.

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